Abstract
Acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT; EC 2.3.1.23) catalyzes the acyl-CoA-dependent acylation of lysophosphatidylcholine (LPC) to produce PC and CoA. LPCAT activity may affect the incorporation of fatty acyl moieties at the sn-2 position of PC where PUFA are formed and may indirectly influence seed TAG composition. LPCAT activity in microsomes prepared from microspore-derived cell suspension cultures of oilseed rape (Brassica napus L. cv Jet Neuf) was assayed using [1-14C]acyl-CoA as the fatty acyl donor. LPCAT activity was optimal at neutral pH and 35°C, and was inhibited by 50% at a BSA concentration of 3 mg mL−1. At acyl-CoA concentrations above 20 μM, LPCAT activity was more specific for oleoyl (18∶1)-CoA than stearoyl (18∶0)- and palmitoyl (16∶0)-CoA. Lauroyl (12∶0)-CoA, however, was not an effective acyl donor. LPC species containing 12∶0, 16∶0, 18∶0, or 18∶1 as the fatty acyl moiety all served as effective acyl acceptors for LPCAT, although 12∶0-LPC was somewhat less effective as a substrate at lower concentrations. The failure of LPCAT to catalyze the incorporation of a 12∶0 moiety from acyl-CoA into PC is consistent with the tendency of acyltransferases to discriminate against incorporation of this fatty acyl moiety at the sn-2 position of TAG from the seed oil of transgenic B. napus expressing a medium-chain thioesterase.
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Abbreviations
- 12∶0:
-
lauroyl
- 16∶0:
-
palmitoyl
- 18∶0:
-
stearoyl
- 18∶1:
-
oleoyl
- 22∶1:
-
erucoyl
- ACBP:
-
acyl-CoA binding protein
- LPAAT:
-
acyl-CoA:lysophosphatidate acyltransferase
- LPCAT:
-
acyl-CoA:lysophosphatidylcholine acyltransferase
References
Stymne, S., and Stobart, A.K. (1987) Triacylglycerol Biosynthesis, in The Biochemistry of Plants-Lipids: Structure and Function (Stumpf, P.K., ed.), Vol. 9, pp. 175–214, Academic Press, New York.
Weselake, R.J., and Taylor, D.C. (1999) The Study of Storage Lipid Biosynthesis Using Microspore-Derived Cultures of Oilseed Rape, Prog. Lipid Res. 38, 401–460.
Weselake, R.J. (2000) Lipid Biosynthesis in Cultures of Oilseed Rape, In Vitro Cell. Dev. Biol.—Plant 36, 338–348.
Weselake, R.J. (2002) Biochemistry and Biotechnology of Triacylglycerol Accumulation in Plants, in Lipid Biotechnology (Kuo, T.M., and Gardner, H.W., eds.), pp. 27–56, Marcel Dekker, New York.
Cao, Y.-Z., and Oo, K.-C. (1990) Lysophosphatidate Acyltransferase in the Microsomes from Maturing Seeds of Meadowfoam (Limnanthes alba), Plant Physiol. 94, 1199–1206.
Bernerth, R., and Frentzen, M. (1990) Utilization of Erucoyl-CoA by Acyltransferases from Developing Seeds of Brassica napus (L.) Involved in Triacylglycerol Biosynthesis, Plant Sci. 67, 21–28.
Oo, K.-C., and Huang, A.H.C. (1989) Lysophosphatidate Acyltransferase Activities in the Microsomes from Palm Endosperm. Maize Scutellum, and Rapeseed Cotyledon of Maturing Seeds, Plant Physiol. 91, 1288–1295.
Voelker, T.A., Hayes, T.R., Cranmer, A.C., and Davies, H.M. (1996) Genetic Engineering of a Quantitative Trait: Metabolic and Genetic Parmeters Influencing the Accumulation of Laurate in Rapeseed, Plant J. 9, 229–241.
Wiberg, E., Edwards, P., Byrne, J., Stymne, S., and Dehesh, K. (2000) The Distribution of Caprylate, Caprate and Laurate in Lipids from Developing and Mature Seeds of Transgenic Brassica napus L., Planta 212, 33–40.
Knutzon, D.S., Hayes, T.R., Wyrick, A., Xiong, H., Davies, H.M., and Voelker, T.A. (1999) Lysophosphatidic Acid Acyltransferase from Coconut Endosperm Mediates the Insertion of Laurate at the sn-2 Position of Triacylglycerols in Lauric Rapeseed Oil and Can Increase Total Laurate Levels, Plant Physiol. 120, 739–746.
Lands, W.E.M., and Hart, P. (1965) Metabolism of Glycerolipids. VI. Specificities of Acyl Coenzyme A:Phospholipid Acyltransferases, J. Biol. Chem. 240, 1905–1911.
Slack, C.R., Campbell, E.C., Browse, J.A., and Roughan, P.G. (1983) Some Evidence for the Reversibility of the Cholinephosphotransferase-Catalysed Reaction in Developing Linseed Cotyledons in vivo, Biochim. Biophys. Acta 754, 10–20.
Slack, C.R., Roughan, P.G., Browse, J.A., and Gardiner, S.E. (1985) Some Properties of Cholinephosphotransferase from Developing Safflower Cotyledons, Biochim. Biophys. Acta 833, 438–448.
Stobart, A.K., and Stymne, S. (1985) The Interconversion of Diacylglycerol and Phosphatidylcholine During Triacylglycerol Production in Microsomal Preparations of Developing Cotyledons of Safflower (Carthamus tinctorius L.), Biochem. J. 232, 217–221.
Vogel, G., and Browse, J. (1996) Cholinephosphotransferase and Diacylglcyerol Acyltransferase. Substrate Specificities at a Key Branch Point in Seed Lipid Metabolism, Plant Physiol. 110, 923–931.
Dahlqvist, A., St⇘hl, U., Lenman, M., Banas, A., Lee, M., Sandager, L., Ronne, H., and Stymne, S. (2000) Phospholipid:Diacylglycerol Acyltransferase: An Enzyme That Catalyzes the Acyl CoA-Independent Formation of Triacylglycerol in Yeast and Plants, Proc. Natl. Acad. Sci. USA 97, 6487–6492.
Stymne, S., and Glad, G. (1981) Acyl Exchange Between Oleoyl-CoA and Phosphatidylcholine in Microsomes of Developing Soya Bean Cotyledons and Its Role in Fatty Acid Desaturation, Lipids 16, 298–305.
Stymne, S., Stobart, A.K., and Glad, G. (1983) The Role of the Acyl-CoA Pool in the Synthesis of Polyunsaturated 18-Carbon Fatty Acids and Triacylglycerol Production in the Microsomes of Developing Safflower Seeds, Biochim. Biophys. Acta 752, 198–208.
Stymne, S., and Stobart, A.K. (1984) Evidence for the Reversibility of the Acyl-CoA:Lysophosphatidylcholine Acyltransferase in Microsomal Preparations from Developing Safflower (Carthamus tinctorius L.) Cotyledons and Rat Liver, Biochem. J. 223, 305–314.
Stymne, S., and Stobart, K. (1986) The Effect of Temperature on the Activity of 2-Oleoyl-sn-phosphatidylcholine Desaturase in Modified Microsomal Membranes from the Cotyledons of Maturating Safflower Seed, Physiol. Vég. 24, 45–51.
Banaś, A., Johansson, I., and Stymne, S. (1992) Plant Microsomal Phospholipases Exhibit Preference for Phosphatidylcholine with Oxygenated Acyl Groups, Plant Sci. 84, 137–144.
Lin, J.T., Woodruff, C.L., Lagouche, O.J., McKeon, T.A., Stafford, A.E., Goodrich-Tanrikulu, M., Singleton, J.A., and Haney, C.A. (1998) Biosynthesis of Triacylglycerols Containing Ricinoleate in Castor Microsomes Using 1-Acyl-2-oleoyl-sn-glycero-3-phosphocholine as the Substrate of Oleoyl-12-hydroxylase, Lipids 33, 59–69.
Moreau, R.A., and Stumpf, P.K. (1982) Solubilization and Characterization of an Acyl-Coenzyme A O-Lysophospholipid Acyltransferase from the Microsomes of Developing Safflower Seeds, Plant Physiol. 69, 1293–1297.
Ichihara, K., Mae, K., Sano, Y., and Tanaka, K. (1995) 1-Acylglycerophosphocholine O-Acyltransferase in Maturing Safflower Seeds, Planta 196, 551–557.
Taylor, D.C., Weber, N., Hogge, L.R., and Underhill, E.W. (1990) A Simple Enzymatic Method for the Preparation of Radiolabeled Erucoyl-CoA and Other Long-Chain Fatty Acyl-CoAs and their Characterization by Mass Spectrometry, Anal. Biochem. 184, 311–316.
Orr, W., Keller, W.A., and Singh, J. (1986) Induction of Freezing Tolerance in an Embryogenic Cell Suspension Culture of Brassica napus by Abscisic Acid at Room Temperature, J. Plant Physiol. 126, 23–32.
Choy, P.C., Tardi, P.G., and Mukherjee, J.J. (1992) Lysophosphatidylcholine Acyltransferase, Methods Enzymol. 209, 80–86.
Bradford, M.M. (1976) A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding, Anal. Biochem. 72, 248–254.
Little, D., Weselake, R., Pomeroy, K., Furukawa-Stoffer, T., and Bagu, J. (1994) Solubilization and Characterization of Diacylglycerol Acyltransferase from Microspore-Derived Cultures of Oilseed Rape, Biochem. J. 304, 951–958.
Bafor, M., Stobart, K., and Stymne, S. (1990) Properties of Glycerol Acylating Enzymes in Microsomal Preparations from Developing Seeds of Safflower (Carthamus tinctorius) and Turnip Rape (Brassica campestris) and Their Ability to Assemble Cocoa-Butter Type Fats, J. Am. Oil Chem. Soc. 67, 217–225.
Larson, T.R., and Graham, I.A. (2001) A Novel Technique for the Sensitive Quantification of Acyl CoA Esters from Plant Tissues, Plant J. 25, 115–125.
Tocher, D.R., Leaver, M.J., and Hodgson, P.A. (1998) Recent Advances in the Biochemistry and Molecular Biology of Fatty Acyl Desaturases, Prog. Lipid Res. 37, 73–117.
Hills, M.J., Dann, R., Lydiate, D., and Sharpe, A. (1994) Molecular Cloning of a cDNA from Brassica napus L. for a Homologue of Acyl-CoA-Binding Protein, Plant Mol. Biol. 25, 917–920.
Engeseth, N.J., Pacovsky, R.S., Newman, T., and Ohlrogge, J.B. (1996) Characterization of an Acyl-CoA-Binding Protein from Arabidopsis thaliana, Arch. Biochem. Biophys. 331, 55–62.
Brown, A.P., Johnson, P., Rawsthorne, S., and Hills, M.J. (1998) Expression and Properties of Acyl-CoA Binding Protein from Brassica napus, Plant Physiol. 26, 629–635.
Chye, M.-L. (1998) Arabidopsis cDNA Encoding a Membrane-Associated Protein with an Acyl-CoA Binding Domain, Plant Mol. Biol. 38, 827–838.
Chye, M.-L., Li, H.-Y., and Yung, M.-H. (2000) Single Amino Acid Substitutions at the Acyl-CoA-Binding Domain Interrupt [14C]Palmitoyl-CoA Binding of ACBP2, an Arabidopsis Acyl-CoA-Binding Protein with Ankyrin Repeats, Plant Mol. Biol. 44, 711–721.
Farooqui, A.A., Horrocks, L.A., and Farooqui, T. (2000) Deacylation and Reacylation of Neural Membrane Glycerophospholipids, J. Mol. Neurosci. 14, 123–135.
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Furukawa-Stoffer, T.L., Boyle, R.M., Thomson, A.L. et al. Properties of lysophosphatidylcholine acyltransferase from Brassica napus cultures. Lipids 38, 651–656 (2003). https://doi.org/10.1007/s11745-003-1110-0
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DOI: https://doi.org/10.1007/s11745-003-1110-0